During the coming year we plan to further study N- and O-glycosylation of hCG synthesized in BeWo cells. In particular we will focus on the effects of an uncoupler, CCCP, and an ionophore, monesium, on the glycosylation process and on secretion. The former drug is believed to block migration of secretory products form the RER to the Golgi, while the latter only affects migration from the Golgi to the plasma membrane. Thus if the predicted sequence of events in N- and O-glycosylation is correct, these drugs should have very different effects on N- and O-glycosylation. Additionally, we plan to use LM cells in culture to study regulation of synthesis of dolichol-PP- oligosaccharide.